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Applied and Environmental Microbiology, June 2000, p. 2541-2547, Vol. 66, No. 6
Department of Plant Pathology, University of
Wisconsin-Madison, Madison, Wisconsin1;
ARIAD Pharmaceuticals, Inc., Cambridge,
Massachusetts2; and Department of
Chemistry and Chemical Biology, Cornell University, Ithaca, New
York3
Received 29 November 1999/Accepted 23 March 2000
Recent progress in molecular microbial ecology has revealed that
traditional culturing methods fail to represent the scope of microbial
diversity in nature, since only a small proportion of viable
microorganisms in a sample are recovered by culturing techniques. To
develop methods to investigate the full extent of microbial diversity,
we used a bacterial artificial chromosome (BAC) vector to construct
libraries of genomic DNA isolated directly from soil (termed
metagenomic libraries). To date, we have constructed two such
libraries, which contain more than 1 Gbp of DNA. Phylogenetic analysis
of 16S rRNA gene sequences recovered from one of the libraries
indicates that the BAC libraries contain DNA from a wide diversity of
microbial phyla, including sequences from diverse taxa such as the
low-G+C, gram-positive Acidobacterium,
Cytophagales, and Proteobacteria. Initial
screening of the libraries in Escherichia coli identified
several clones that express heterologous genes from the inserts,
confirming that the BAC vector can be used to maintain, express, and
analyze environmental DNA. The phenotypes expressed by these clones
include antibacterial, lipase, amylase, nuclease, and hemolytic
activities. Metagenomic libraries are a powerful tool for exploring
soil microbial diversity, providing access to the genetic information
of uncultured soil microorganisms. Such libraries will be the basis of
new initiatives to conduct genomic studies that link phylogenetic and
functional information about the microbiota of environments dominated
by microorganisms that are refractory to cultivation.
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Cloning the Soil Metagenome: a Strategy for Accessing the Genetic
and Functional Diversity of Uncultured Microorganisms
*
Corresponding author. Mailing address: Department of
Plant Pathology, University of Wisconsin-Madison, 1630 Linden Drive, Madison, WI 53706. Phone: (608) 263-8783. Fax: (608) 262-8643. E-mail:
joh{at}plantpath.wisc.edu.
Present address: Vertex Pharmaceuticals, Inc., Cambridge, MA 02139.
Present address: UCB Research Inc., Cambridge, MA 02139.
§
Present address: Biogen, Inc., Cambridge, MA 02142.
Present address: The Ohio State University, Columbus, OH 43210.
Applied and Environmental Microbiology, June 2000, p. 2541-2547, Vol. 66, No. 6
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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